Project Summary Yale Cooperative Center of Excellence in Hematology Imaging Core The Imaging Core of the YCCEH has provided access to state-of-the-art microscopy for benign hematology to progressively more users each year. In this renewal, we propose to continue operations with enhancements including longterm timelapse with decreased phototoxicity, adapted super resolution imaging for hematopoietic tissues and automated high-throughput microscopy for imaging of 1,000 to 10,000 cells/24 hrs at super- resolution. Hematopoiesis is a complex and dynamic process involving many heterologous cell types. Due to the rich spatial and/or temporal information contained in imaging data, imaging is uniquely powerful in revealing the influences of cellular dynamics and tissue organization on the relevant cells and states. As many hematopoietic cells are small in size, minimally adhesive, difficult to manipulate genetically, reside in deep tissues, highly dynamic and/or exceedingly rare, the ability to apply imaging approaches to hematopoiesis research has been limited to few laboratories with the requisite technical expertise and instrumentation. YCCEH Imaging Core aims to reduce the high technical hurdle and promote the application of imaging techniques for hematopoietic research. Yale is the home of many state-of-the-art imaging equipment and facilities overseen by world-renowned leaders and experts in microscopy. The YCCEH Imaging Core will make the large array of imaging platforms readily accessible to all investigators with a research interest in hematopoiesis, locally or nationally. YCCEH Imaging Core will continue to provide its members with access to state-of-the-art instrumentation including widefield, confocal, 2-photon and flow cytometry imaging. Importantly, through this Imaging Core, we are making it available as a service, access to and expertise in using innovative imaging modalities including automated long-term live-cell time-lapse microscopy, optical clearing of tissue and super-resolution microscopy. The directors of the Core, Drs. Bewersdorf and Guo will act as liaisons between YCCEH members and the participating facilities and labs and will help identifying most suitable techniques for individual research projects. The Imaging Core will also support two part time imaging specialists who will train YCCEH members, maintain the instruments provided by the Bewersdorf and Guo labs and assist with reagents, sample preparation and data analysis. We will continue to promote the dissemination of knowledge for imaging in hematopoietic research by organizing symposia, workshops, seminars and training with an emphasis on students, trainees and other early-stage investigators. Furthermore, the Imaging Core will continue to develop, optimize and disseminate innovative reporters/probes that are suitable for analyzing hematopoietic cells by fluorescence imaging and/or flow cytometry. We will continue to encourage YCCEH members to evaluate various microscopy techniques for their research projects through the user incentive program, by eliminating the psychological hurdle of investing funds into an unfamiliar technical domain. Overall, the Imaging Core will facilitate and enable individual hematopoietic researchers to make observations of their specific hematopoietic cell linages, using approaches that can provide unique structural, functional and/or dynamic information.